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Originally published as MBC in Press, 10.1091/mbc.E03-04-0214 on June 27, 2003

Vol. 14, Issue 10, 4028-4038, October 2003

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Localization of Pavarotti-KLP in Living Drosophila Embryos Suggests Roles in Reorganizing the Cortical Cytoskeleton during the Mitotic Cycle

Gianluca Minestrini, Alyssa S. Harley, and David M. Glover *

Cancer Research UK, Cell Cycle Genetics Group, University of Cambridge, Department of Genetics, Cambridge CB2 3EH, United Kingdom

Submitted April 8, 2003; Revised May 28, 2003; Accepted May 28, 2003
Monitoring Editor: J. Richard McIntosh

Pav-KLP is the Drosophila member of the MKLP1 family essential for cytokinesis. In the syncytial blastoderm embryo, GFP-Pav-KLP cyclically associates with astral, spindle, and midzone microtubules and also to actomyosin pseudocleavage furrows. As the embryo cellularizes, GFP-Pav-KLP also localizes to the leading edge of the furrows that form cells. In mononucleate cells, nuclear localization of GFP-Pav-KLP is mediated through NLS elements in its C-terminal domain. Mutants in these elements that delocalize Pav-KLP to the cytoplasm in interphase do not affect cell division. In mitotic cells, one population of wild-type GFP-Pav-KLP associates with the spindle and concentrates in the midzone at anaphase B. A second is at the cell cortex on mitotic entry and later concentrates in the region of the cleavage furrow. An ATP binding mutant does not localize to the cortex and spindle midzone but accumulates on spindle pole microtubules to which actin is recruited. This leads either to failure of the cleavage furrow to form or later defects in which daughter cells remain connected by a microtubule bridge. Together, this suggests Pav-KLP transports elements of the actomyosin cytoskeleton to plus ends of astral microtubules in the equatorial region of the cell to permit cleavage ring formation.


Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03–04–0214. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03-04-0214.

Online version of this article contains video material. Online version is available at www.molbiolcell.org.

* Corresponding author. E-mail address: dmg25{at}mole.bio.cam.ac.uk.




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